Note: Descriptions are shown in the official language in which they were submitted.
W093/~02 - PCT/US92/07622
21189~9
BIOLOGICALLY ACTIVE AMPHIPHILIC PEPTIDE ~;
COMPOSITIONS AND USES THEREFOR
This application is a contin~ation-in-part of Applica-
tion Serial No. 760,054, filed September 13, 1991.
Thi8 invention relates to biologically active
amphiphilic peptides. More particularly, thi~ invention .-.
relates to biologically active amphiphilic peptides u~eful
in pharmaceutical compositions.
In accordance with an aspect of the pre~ent invention,
there i~ provided a biologically active amphiphilic peptide
which ~ncludes the following basic ~tructure X~O: .
Rl-R2-R2-Rl-R2-R2-Rl Rl R2 ,1
Rl is a hydrophobic amino acid, and R2 is a basic
hydrophilic or neutral hydrophilic amino acid.
In one embodiment, the peptide includes the basic
~tructure Y10-X10 wherein X10 is as hereinabove described
and Yl~ is
(i~ Rl;
(ii) R2-Rl; or
~ iii) R2-R2-Rl, wherein Rl and R2 are as hereinabove~
de~cribed.
The hydrophobic amino may be selected from the cla~s
consi~ting of Ala, Cys, Phe, Gly, Tle, Leu, Met, Pro, Val,
Trp and Tyr.
SUE3SllTUJTE SHEEr
WO g3/05802 PCr/USg2/07622
2118939 -2-
The basic hydrophilic amino acid acids may be selected
from the clas~ consi~ting of Lys, Arg, Hi6, Orn,
homoarginine (Har), 2, 4-diaminobutyric acid (Dbu), and
p-aminophenylalanine. The neutral hydrophilic amino acids
may be selected from the class consisting of A~n, Gln, Ser,
Thr, and homoserine (H~e).
In one embodiment, Rl i~ leucine. In another embodiment, R2
is lysine. Representative examples of peptide~ in accor-
dance with thi~ aspect of the present invention include
those having the following structures:
(SEQ ID NO: 1)
ISEQ ID NO: 2)
(SEQ ID NO: 3)
(SEQ ID NO: 4) ~
In accordance with another aspect of the present -`
invention, there is provided a biologically active
amphiphilic peptide which includes the following basic
~tructure X12:
. .
2 1 2 R2 Rl Rl R2-R2-Rl-R2-R2, wherein Rl i8 a
hydrophobic amino acid and R2 i~ a baslc hydrophilic or
neutral hydrophilic amino acid.
- In:one embodiment Rl i8 leucine. In another embodi-
ment, R2 i 8 ly~ine.
In one embodiment, the peptide include~ the basic
structure Y~2-X12, wherein X12 i~ as hereinabove described,
and Y12 i~
(i) R2;
(ii) Rl-R2;
('iii) 1 Rl-R2;
(iv) R2 Rl R1 R2; or
(V) ` R2-R2-Rl-Rl R2-
In one embodiment, the peptide may have the following
structure; -~
: ~ SUBSrlTUTE SHEET
:~:
~ -
.~ .
W093/~2 2118 9 3 ~CT/US92/07G22
Lys Ly~ Leu Leu Lys Lys Leu Ly~ Lys Leu
Leu Ly~ Ly~ Leu Arg Arg
(SEQ ID NO.:5)
In another embodiment, the peptide includes the ba~ic
X12 Z12' wherein X12 is as hereinabove de-
scribed, and Z12 i8:
(i) R~
(ii) Rl-Rl;
(iii) Rl-Rl-R2;
(iv) Rl R1 R2 R2; or
( v ) Rl-Rl -R2-R2 Rl i
In one embodiment, the peptide may have the following
structure:
Lys Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys
Leu
5 ~0 15
(SEQ ID NO:6)
In another embodiment, the peptide may include ~he
&tructure:
(y ) - X12 ~ (Zi2)b' wherein X12' Y12 a 12
hereinabove described, and a is 0 or 1, and b iB O or 1.
In accordance with another a~pect of the pre~ent
. . in~e~tion, there is provided a biologically a~tive
amphiphilic peptide which include~ the following ba~ic
structure X14:
1 2 2 Rl Rl R2 R2-Rl-R2-R2-R1 Rl-R2_R -R
wherein R$ and R2 are a~ hereinabove described, and R3 is a
neutral hydrophilic amino acid.
In one embodiment, the peptide may have the following
structure:
(SEQ ID NO:7)
SUBSTITUTE SI~E~
rcr/uss2/07622 :~
W093/0~2 ; i .
~118g39
In another embodiment, the peptide may have the follow- .
ing structure: ;~
(SEQ ID N0:8
In accordance with yet another aspect of the present
invention, there i8 provided a biologically active
amphiphilic peptide which includes the following basic
structure X16:
1 2 1 1 2 R2 Rl Rl-R2-R2-R4, wherein Rl and R2 are as
hereinabove de~cribed, and R4 i8 a neutral hydrophilic amino .`
acid or proline.
In one embodiment, the peptide may include the follow-
ing 8tructure Y16-X16) wherein X16 is the basic peptide
structure hereinabove de~cribed, and Y16 i8:
(i) -Rl; ,,
(ii) -Rl-Rl; :
(iii) -R2-Rl-Rl;
(iv) -Rl~R2~Rl Rl;
(v) -Rl-Rl-R2-Rl Rl;
(vi) -R2-Rl-Rl-R2-Rl Rl;
(Vii ) -R2-R2-Rl-Rl-R2 Rl Rl,
wherein Rl and R2 are as herelnabove described.
In one embodiment, the peptide may include the struc-
ture-:
X16-Zl6, wherein X16 is as hereinabove described, and
Z16 i8:
(i) -R2;
( ~1 i ) -R2-R2;
(iii) -R2-R2 Rl;
(iv) -R2~R2~Rl Rl;
(v) -R2-R2-Rl-Rl-R2;
(vi) -R2-R2-Rl-Rl R2 R2;
(Vii ) -R2-R2-Rl-Rl-R2 R2 Rl
In a preferred embodiment, the peptide may have one of
the following structures:
~ ~ .
; :~
~ ~ .
SlJ8Srml~ SHE~
,,,
" .
W093/~2 rCT/US92/07622
2118939
(SEQ ID NO:9); or
(SEQ ID NO:lO).
In another embodiment, the peptide may have the struc-
tUre (Y ) -Xl6-~Zl6)b~ wherein Xl6, Yl6, 16
hereinabove described, a i~ O or l, and b i6 0 or l.
In accordance with another a~pect of the present
invention, there i~ provided a biologically active
amphiphilic peptide which include6 the following basic
structure Xl8:
1 R2 R2 Rl-R2-R2-Rl-Rl-R2-R2-Rl-R3
wherein Rl, R2, and R3 are as hereinabove described.
In accordance with another embodiment, the peptide may
include the structure Yl8-Xl8, wherein X18 i8 as hereinabove
described, and Yl8 i~:
(i) -Rl; ;~
(li) -R2-Rl; .;
(iii) -R2-R2-Rl;
iv~ -Rl-R2-R2 Rl;
(v)~ -Rl-Rl-R2-R2 Rl;
(vi) -R2-Rl-Rl-R2-R2 Rl;
2 2 Rl Rl ~ -R2-Rl, wherein Rl and R are as
hereinabove described.
In another embodiment, the peptide includes the struc-
: ture xl8-æl8~ wherein Xl8 i6 as hereinabove described, ana
Z18 i8:
Rl;
(ii) -Rl-R5;
(iii) -Rl-R5-R5;
(iv) -Rl-R5-R5-R3;
(v) -Rl-R5~RS~R3 Rl;
(viJ -Rl-R5-R5-R3 Rl R3;
(vll) -Rl-R5-Rs-R3-Rl R3 R3;
viii) -Rl-Rs-Rs-R3-Rl-R3 R3 R5;
' :
, ;, ~ ~,
~,~. ., : ~ :
. S~E~S~ITIJ~E SHEEr
.~ .,
- W093/0~2 PCT/US92/07622
-6-
1 5 5 3 Rl R3 R3-R5-R3, wherein Rl and R are
as hereinabove described, and R5 is proline. :~
In one embodiment, the peptide has the following
structure: ::
(SE~ ID N0~
In one embodiment, the peptide may have the structure ::
X18 (Zi8)b~ wherein X18' Y18' and Z18 are as
hereinabove described, a i8 0 or 1, and b i~ 0 or 1. ~
In accordance with another aspect of the present ~;
invention, there is provided a biologically active -~
amphiphilic peptide which includes the following basic ~:~
structure X20:
1 1 3 2 1 1 Rl Rl Rl-Rl-R2-Rl-Rl-R2-R2-Rl-Rl-R -R -R
_,
wherein Rl, R2, and R3 are as hereinabove described. In one
-.:
embodiment, the peptide may have the following structure:
(SEQ ID N0:12). ~:
In another`embodiment, the peptide may include the ~:
structure X20-Z20, wherein X20 i8 as hereinabove described, --
and Z20 i~
(i) -R2;
( ii ) -R2-R2; ~;
(iii) -R2-R2-R~
(iv) -R2-R2-Rl Rl;
(v) -R2-R2-Rl Rl R2;
(vi) -R2-R2-R1-Rl R2 ~2;
(Vii ) -R2-R2-Rl-Rl R2 R2 Rl
In accordance with yet another aspect of the present
invention, there is provided a biologically active
amphiphilic peptide having a structure selected from the
group consi~ting of
(a) R1-R2-R2-Rl R2 R2 Rl;
(b) Rl-Rl-R2-R2-Rl R2 R2 Rl;
; ( ) R2 Rl Rl-R2-R2-Rl-R2-R2-Rl;
..
'?~ .
,': '~ . ,
... .
SUBS~ITUTE S!~E~,
W093/0~2 PCT/US92/07622
7_
2118939
(d) R2-R2-Rl-Rl-R2-R2-Rl-R2 R2 Rl;
1 2 R2 Rl Rl R2-R2-Rl-R2-R2-Rl~ wherein Rl and
R2 are as hereinabove deseribed.
In one embodiment, the peptide ha~ the structure ~a),
and a representative example of such a structure is (SEQ ID
N0:13), whieh is given in the accompanying sequenee ligting.
In another embodiment, the peptide has the structure
(b), and a representative example of such a ~tructure i~
(SEQ ID N0:14), whieh i~ given in the accompanying sequence
listing.
In another embodiment, the peptide ha~ the structure
(e), and a repre~entative example of sueh a structure is
(SEQ ID N0:15) as given in the aeeompanying ~equenee list-
. ing.
In yet another embodiment, the peptide has the struc-
ture (d), a~d a repre~entative example of sueh a strueture
i~ (SEQ ID N0:16) as given in the aeeompanying ~eguenee
listing.
In a further embod~me~t, the peptide ha# the strueture
(e), and representative examples of sueh a ~trueture are
(SEQ ID N0:17) and (SEQ ID N0:18) as given in the aeeompany-
~- ing ~egu~nee listing.
~- In aeeordanee with another aspeet of the present
invention, there is provided a biologically active
amphiphilie peptide having the following struetural formula:
(SEQ ID N0:19).
In general, the peptides hereinabove described are ion
ehannel-forming peptides.
- An ion ehannel-forming peptide or ionophore is a
peptide whieh inerea~es the permeability for ions aeross a
natural or synthetic lipid membrane. B. Christen~en et
al., PNAS, Vol. 85 Pgs. 5072-76 (July, 1988) and Anzai, et
al., Bioehimiea et Bio~h~siea Aeta., Vol. 1064, pgs. 256-266
(1991), deseribe methodology whieh indieates whether or not
~, : , . .
~- .
,.. .
, :
'~' ~ . ' f
SUBSTITU~E SHEET
:
W093/05802 PCT/US92/07622
-8-
893g
a peptide has ion channel-forming properties and is there-
fore an ionophore. As u ed herein an ion channel-forming
peptide is a peptide which has ion channel-forming proper-
ties as determined by the method of Christensen et al or
Anzai, et al.
In one embodiment, the hereinabove de~cribad peptide~
may be acetylated with a CH3C0-group at the N-terminal, said
CH3CO-group being hereinafter referred to by the letter X.
In another embodiment, the hereinabove described
peptides may include an octanoyl group at the N-terminal.
In accordance with one embodiment, each of the amino
acid residues contained in the peptides which i8 not
glycine, i8 a D-amino acid residue. Although the ~cope of
thi# particular embodiment i~ not to be limited to any
theoretical rea~oning, it is believed that the above-men-
tioned peptide~, when con~isting entirely of D-amino acid or
glycine re~id~es, may have increa~ed re~i~tance to
proteolytic enzymes while retaining their biol~gical activi-
ty. Such peptide~ th~8 may be administered oralIy. Al~o,
in accordance with another embodiment, all of the amino aci~
residue~ may be D-amino acid or glycine re~idue~, or L-amino
acid or glycine re id~e~.
In ~eneral, the peptides and/or analogues or deriva-
tives thereof are generally water ~oluble to a concentration
of at lea~t 20 mg/ml at neutral pH in water. In addition,
the structure of such peptide provides for 1exibility of
the peptide molecule. When the peptide is pla~ed in water,
it doe~ not a~ume an amphiphilic structure. When the
peptide encounter~ an oily surface or membrane, the peptide
chain fold~ upon it~elf into a rod-like ~tructure.
SlJBsTlTlJTE S~T
W093/0~2 PCT/US92/07622
_g ; ~
21I8939
The peptides and/or analogues or derivatives thereof
may be administered to a host; for example a human or
non-human animal, in an amount effective to inhibit growth
of a target cell, virus, or virally-infected cell. Thus,
for example, the peptides and/or analogues or derivatives
thereof may be u~ed as antimicrobial agents anti-viral
agent~, anti-bacterial àgents, anti-tumor agents, anti-para-
sitic agents, spermicide~, as well as exhibiting other ~-
bioactive functions. r
The term "antimicrobial" as used herein mean~ that the
polypeptides of the present invention inhibit, prevent, or
destroy the growth or prolieration of microbes such as
bacteria, fungi, viruses, or the like.
The term "anti-bacterial" as used herein mean~ that the
polypeptide~ employed in the present invention produce
effects adverse to the normal biological functions of
bacteria, lncluding death or destruction and prevention of
the growth or proliferation of the bacteria when contacted
with the po:lypeptide~.
The t-rm "antibiotic" as used herein means that the
peptide~ employed in the pre~ent lnvention produce effect~
advers- to the normal biological functions of~he non-host
cell, tissue or organism, including death or destruction and
prevention of the growth or proliferation of the non-host '
cell, ti#sue, or organism when contacted with the peptides.
The term "~permicidaln as u~ed herein means that the
polypoptides employed in the pre~ent invention, inhibit,
prevent, or de~troy the motility of sperm.
The term "antiviral" as used herein mean~ that the
polypepSide~ employed in the pre~ent invention inhibit,
prevent, or destroy the growth or proliferation of viru~e~,
or of virally-infected cell~.
.~, ~ , .
,.. . .
~ SUBSmUTE SHEEr
W093/0~2 - PCT/US92/07622
211~93~ o-
The term "anti-tumor" a~ used herein mean~ that the
polypeptide inhibit~ the growth of or destroys tumors,
including cancerous tumorg.
The term "anti-parasitic" as used herein means that the
polypeptides employed in the present invention inhibit,
prevent, or destroy the growth or proliferation of para-
sites.
The peptides of the present invention have a broad
ranae of potent antibiotic activity against a plurality of
microorganisms including gram-positive and gram-negative
bacteria, fungi, protozoa, and the like, as well as para-
sites. The peptides of the pre~ent invention allow a method
for treating or controlling microbial infection cauQed by
organisms which are ~ensitive to the peptides. Such treat-
ment may comprise admini~tering to a host organism or tissue
susceptible to or affiliated with a microbial infection an
antimicrobial amount of at lea~t one of the peptideQ.
Becau~e of the antibiotic, antimicrobial, antiviral,
and antiba~terial properties of the peptide~, they may also
be uQed as pre~ervatives or sterilants or disinfectant~ of
materials ~u~ceptible to microbial or viral contamination.
The peptides and/or derivatives or analogues thereof
may be admini~tered in combination with a non-toxic pharma-
ceutical carrier or vehicle ~uch as a filler, non-toxic
buffer, or physiological saline solution. Such pharmaceuti-
cal compositions may be used topically or systemically and
may be in any suitable form ~uch as a liguid, solid, semi-
solid, injectable ~olution, tablet, ointment, lotion, paste,
- capsule, or the like. The peptide compo~ition~ may also be
used in combination with adjuvant~, protea~e inhi~itor~, or
compatible drugs where ~uch a combination is ~een to be
d~sirable or advantageou~ in controlling infection cau~ed by
harmful microorganisms including protozoa, viru~es, and the
like, as well aQ by parasites.
~ JBS~lT~JTE S~E
`
W093/0~2 PCT/USg2/0762~
2118~39
The peptides of the pre~ent invention may be admini~-
tered to a host; in particular a human or non-human animal,
in an effective antibiotic and/or anti-tumor and/or anti-vi-
ral and/or anti-microbial and/or antibacterial and/or
anti-parasitic and/or an antispermicidal amount.
Depending on the u~e, a composition in accordance with
the invention will contain an effective anti-microbial
amount and/or an effective antispermicidal amount and/or an
effective anti-viral amount and/or an effective anti-tumor
amount and/or an effective anti-parasitic and/or an effec-
tive antibiotic amount of one or more of the hereinabove
described peptides which have such activity. The peptides
may be administered by direct application of the peptides to
the target cell or virus or virally-infected cell, or
indirectly applied through ~ystemic admini~tration.
The peptides of the present invention may also be
employod in promoting or ~timulating healing of a wound in a
host.
The term "wound healing" as used herein includes
various aspects of the wound healing proces6.
These aspect~ include, but are limited to, increased
contrac~ion of the wound, increased depo~ition of connective
tis~ue, as evidenced by, for example, increa~ed deposition
of collagen in the wound, and increased tensile strength of
the wound, i.e., the peptides increa~e wound breaking
strength. The peptides of the present invention may al80 be
employed 80 as to reverse the inhibition of wound healing
caused by conditions which depress or compromise the immune
system.
The peptides of the pre~ent invention may be u~ed in
the treatment of external burns and to treat and/or prevent
~kin and burn infections. In particular, the peptides may
be uJed to treat ~kin and burn infections cauJed by
,~ ,
SUBS~ rE SHE~
W093/0~2 ~ PCT/US92/07622
-12-
2118939
organisms such a~, but not limited to, P. aeruqinosa and S.
aureus
The peptides are also useful in the prevention or
treatment of eye infections. Such infections may be caused
by bacteria such as, but not limited to, P. aeruainosa, S.
aureus, and N. gonorrhoeae, by fungi such as but not limited
to C. albicans and A. fumiaatus, by parasites such as but
not limited to A. castellani, or by viru~es.
The peptides may also be effective in killing cysts,
spores, or trophozoites of infection - causing organism6.
Such organism~ include, but are not limited to Acanthamoeba
which forms trophozoites or cysts, C. albicans, which forms
spore~, and A. fumiaatu~, which forms spore~ as well.
The peptides may also be administered to plant~ in an
effective antimicrobial or antiviral or antiparasitic amount
to prevent or treat microbial or viral or paras~tic contami-
nation thereof.
The peptides, when u#ed in topical compositions, are
generally present in an amount of at least o.lX, by weight.
~ In most cases, it is not nece~sary to employ the peptide in
;~ an amount greater than 2.0X, by weight.
- In employing such compositions systemicalfy
(intramuscular, intravenous, intraperitoneal), the active
peptide is present in an amount to achieve a serum level of
the peptide of at ~east about 5 ug/ml. In general, the ~erum
level of peptide need not exceed 500 ug/ml. A preerred
serum level is about lO0 ug/ml. Such serum level# may be
achieved ~y incorporating the peptide in a composition to be
administered sy~temically at a dose of from l to about lO
mg/kg. In general, the peptide( 8 ) need not be admini~tered
at a dose exceeding lO0 mg/kg.
The peptides may be produced by known techniques and
- obtained in ~ubstantially pure form. Eor example, the
peptides may be ~ynth-sized on an automatic peptide
,.
, ' ~ '
.`.~: ~
~ ~ SUBSmUTE SHl~E~
,`, ~
W093/0~2 rCT/USg2/07622
-13- 2118939
~ynthesizer. Journal of the American Chemical societY, Vol.
85, pg~. 2149-54 ~1963). It i~ also possible to produce
such peptide~ by genetic engineering technique~.
Thu8, within the scope of the pre~ent invention there
may be provided DNA which encodes the peptides hereinabove
described. Such DNA may be expressed by means known to
those skil1ed in the art. Thus, it is al~o contemplated
that within the scope of the present invention the peptides
may be administered by administering to a ho~t DNA which
encodes the peptide(s).
In accordance with another embodiment, the peptides of
the present invention may be employed in combination with an
ion having phamacological properties for the purpose~
hereinabove described.
An ion having pharmacological propertie~ is one which
when introduced into a target cell, virus, or virally-in-
fected cell, inhiblts and/or prevents and/or destroys the
growth o the target cell, virus, or virally-infected cell.
Such an ion having pharmacological properti-s i~ one
which in the absence of an ion channel forming peptide is
unable to cross a natural or synthetic lipid m~mbrane; in
particular a cell membrane, in sufficient amount~ to affect
a cell or viru8 adversely.
m e peptide and ion having pharmacological properties
may be administered as a single composition or in separate
composition~; and the single or separate compo~itions may
include additional material~, active# and/or inactive~, in
addition to the peptide and ion having pharmacological
propertie~. A6 representative examples of ions having
pharmacological properties which may be employed, there may
be mentioned fluoride, peroxide, bicarbonate, silver, zinc,
mercury, ar~enic, copper, platinum, antimony, gold, thalll-
um, nickel, selenium! bismuth, and cadmium ions.
,
, ~
i .:,~
. ~ .
,.
; ~ SUF~STrrlJT~ SHEES
,~
W093/~02 PCT/US92/07622
-14-
2 1 18939 The peptide and the ion having pharmacological proper-
ties, whether administered or prepared in a single composi-
tion or in separate compositions, are employed in amount~
effective to inhibit and/or prevent and/or destroy the
growth of the target cell. In effect, the ion potentiates
the action of the peptide, i.e., the amount of ion i8
effective.to reduce the maximum effective concentration of
the peptide or protein for inhibiting growth of a target
cell, viru~, or viral~ly-infected cell.
The ion having pharmacological properties, when used
topically, i~ generally employsd in a concentration of from
0.05% to 2.0%. When used ~ystemically, the ion i8 generally
employed in an amount of from 1 to 10 mg. per kg. of host
weight. Peptide dosages may be within the ranges
hereinabove described.
It is also to be understood that the peptide and ion
having pharmacological properties, may be del~vered or
administe~ed in different forms; for example, the toxic ion
may be ad~inistered orally, whlle the peptide may be admin
istered by TV or IP.
As repre~entative examples of administering the peptide
or protein and ion for topical or local administration, the
peptide could bè admini~tered in an amount of up to about lX
weight to weight and the ion delivered in an amount of about
50mM (about 0.1%). Alternatively, the ion, in the form of a
~alt ~uch a~ ~odium fluoride, could be administered orally
in conjunction with ~ystemic admini~tration of the peptide.
Eor example, the peptide may be administered IV or IP to
achieve a serum dose of 100 micrograms per milliliter ~10
milligrams per kilogram) in conjunction with an oral dose of
ion, in particular, sodium fluoride, of 10 meq per kilogram.
In accordance with another embodiment, the peptides of
the pre~ent invention may be administered to a host in
combination with an antibiotic ~elected from the class
S(JBSTITUTE SHEET
~: ....... ....... ..... . ..
W093/0~2 - PCT/USg2/07622
-15-
2118939
consisting of bacitracin~, gramacidin, polymyxin,
vancomycin, teichoplanin, aminoglycosideE, hydrophobic
antibiotics, penicillins, monobactams, or derivative~ or
analogues thereof.
The bacitracins, gramacidin, polymyxin, vancomycin,
teichoplanin, and derivative~ and analogues thereof, are a
group of polypeptide antibiotics. A preferred bacitracin is
bacitracin A.
Aminoglycoside antibiotics include tobramycin,
kanamycin, amikacin, the gentamicins (e.g., gentamicin C1,
gentamicin C2, gentamicin Cla), netilmicin, and derivative~
and analogue~ thereof. The preferred aminoglycosides are
tobramycin and the gentamicins. The aminoglyco#ide~, and
the bacitracins hereinabove described, tend to be
hydrophilic and water-~oluble.
Ponicillin~ which may be employed include, but are not
limited to benzyl penicillin, ampicillin, methicillin
(dimethoxy~phenyl penicillin), ticaricillin, penicillin V
(phenoxymethyl penicillin), oxacillin, cloxacillin,
dicloxacillin, flucloxacillin, amoxicillin, and
amidinocillin. Preferred penicillins which may be employed
are ben~yl penicillin and ampicillin. A preferred
monobactam which may be employed is aztreonam.
A~ repre~entative example~ of hydrophobic antibiotics
which may be used in the pre~ent invention, there may be
mentioned macrolide~ such as erythromycin, roxythromycin,
clarithromycin, etc.; 9-N-alkyl derivative~ of erythromycin;
midecamycin acetate; azithromycin; fiurithromycin;
rifabutin; rokitamycin; a 6-0-methyl erythromycin A known as
TE-031 (Tai~ho); rifapenti~ne; benzypiperazinyl rifamycin~
~uch as CGP-7040, CGP-5909, CGP-279353 (Ciba-Geigy); an
erythromycin A derivative with a cyclic carbamate fused to
the Cll/C12 position of a macrolide ring known ~8 A-62514
(Abbott); AC-7230 (Toyo Jozo); benzoxazinorifamycin;
S~JBSTITUTE SHEEr
.,-,~ ~
, ~, ~
W093/0~2 - PCT/US92/07622
-16-
2~8939
difficidin; dirithromycin; a 3-N-piperdinomethylzaino methyl
rifamycin SV known a~ FCE-22250 (Farmitalia); M-ll9-a (Kirin
Brewery); a 6-0-methyl-1-4"-0-carbamoyl erythromycin known
as A-63075 (Abbott); 3-formylrifamycin SV-hydrazones with
diazabicycloalkyl ~ide chains such as CGP-27557 and CGP-2986
(Ciba-Geigy); and 16-membered macrolides having a
3-0-alpha-L-cladinosyl moiety, ~uch a~
3-0-alpha-L-cladino~yldeepoxy rosaramicin; tylosins and acyl
demycinosyl tylosins.
In addition to the macrolides hereinabove described,
rifamycin, carbenicillin, and nafcillin may be employed as
well.
Other antibiotics which may be u~ed ~whether'or not
'. hydrophobic) are antibiotics which are SO-S ribosome inhibi-
tors such a~ lincomycin; clindamycin; and chloramphenicol;
etc.; antibi'otics which have a large lipid like lactone
ring, ~uch as mystatin; pimaricin, etc.
The peptide and antibiotic may be adminstered by direct
administration to a target cell or by systemic or topical
admini~tration to a host which include~ the target cell, in
order to prevent, destroy or inhibit the growth of a target
cell. Target cells whose growth may be prevented, inhibit-
ed, or de~troyed by the administration of the peptides and ,
'antibiotic include Gram-positive and Gram-negative bacteria
a~ well as fungal cells.
The antibiotic, such as those hereinabove described, or
- derivative~ or analogue~ thereof, when used topically, i8
generally employed in a concet~ation of about 0.1% to about
loX. When used systemically, the antibiotic or derivative
or analogue thereof is generally employed in an amount of
from 1.25 mg. to about 4~ mg. per kg. of host weight per
day. Peptide dosages may be those as hereinabove described.
As repre~entative exmples of administering the peptide
and antibiotic Eor topical or local administration, the
, ~
. : . .
~" ~ ~ ,
-~ ~ SUBSTITUTESHEEl-
W093/~02 PCT/U~92/07622
-17-
211~93~
peptide could be admnistered in an amo~nt of from about 0.1%
to about 10% weight to weight, and the antibiotic i8 deliv-
ered in an amount of from about 0.1~ to about 10% weight to
weight.
In accordance with another embodiment, the peptides of
the present invention may be administered in combination
with an antipara~itic agent or an antifungal agent.
Antiparasitic agents which may be employed include, but
are not limited to, anti-protozoan agents. Examples of
specific anti-parasitic agent~ which may be employed in-
clude, but are not limited to, pentamidine i~ethionate, and
propamidine isethionate (Brolene).
Anti-fungal agents which may be employed include, but
are not limited to, ketoconazole. It is al80 to be under-
stood that certain anti-parasitic agents, may al80 have
anti-fungal activity, and that certain anti-fungal agents
may have anti-parasitic activity.~~
In accordance with another embodiment, the peptides of
the pre~ent invention may be administered in combination
with an antibiotic which inhibits DNA gyra~e, which i8 an
enzyme involved in the formation of bonds between individual
coiling ~tra~d~ of replicating bacterial DNA. ~Thus, DNA
gyra~e i~ neces~ary for the normal replication of bacterial
DNA, and, therefore, antibiotic~ which inhibit DNA gyra~e
inhibit the normal replication of bacterial DNA.
Example~ of antibiotics which i~hibit DNA gyrase
include nalidixic acid, oxolinic acid, cinoxacin, and
quinolone antibiotics which include ciprofloxa~in,
norfloxacin, ofloxacin, enoxacin, pefloxacin, lomefloxacin,
fleroxacin, tosulfloxacint temafloxacin, and rufloxacin.
In accordance with another embodiment, the peptides of
the pre~ent invention may be administered for the purpo~e
hereinabove described in combination with other biologically
SUBSTITUTE S:~IEET
W093/~ ~2 PCT/US92/07622
2~939 -18- ~
active amphiphilic peptides, or in combination wlth ion
channel-forming proteins.
The present invention will be further described with
respect to the following example; however, the scope of the
invention is not to be limited thereby.
EXAMPLE
Table I, which follows, indicates the Minimal
Inhibitory Concentration (MIC) in ~g/ml of Peptide~ (SEQ ID
N0:1) through (SEQ ID N0:3), (SEQ ID N0:5), (SEQ ID N0:6),
(SEQ ID N0:9) through (SEQ ID N0:11), and (SEQ ID N0:17)
againæt S.aureu~ ætrain ATCC 25923, P. aeruainosa strain
ATCC 27853, and E.coli ATCC strain 25922.
The procedure for the antibacterial aæsay is ba~ed upon
the guidelines of the National Committee for Clinical
Laboratory Standards, Document M7-T2, Volume 8, No. 8, 1988.
Stock solutionæ of peptides (SEQ ID N0: 1) through ~SEQ
ID N0:3), (SEQ ID N0:5), (SEQ ID N0:6), (SEQ ID N0:9)
through (SEQ ID N0:12), and (SEQ ID N0:18) in accordance
with the present invention are prepared at a concentration
of 512 ~g/ml in sterile deionized distilled water and stored
at -70C. Each peptide i~ a C-terminal amide 'and may or may
not be acetylated at the N-Terminus. An acetyl group at the
N-terminus i~ indicated by an X.
The ~tock peptide ~olution i~ diluted in ~erial dilu-
tions (1:2) down the well~ of a microtiter plate so that the
final concentrations of peptides in the wellæ are 0.25,
0.50, 1, 2, 4, 8, 16, 32, 64, 128, and 256 ~g/ml. 1-5 X 105
CFU~/ml of either S. aureus ATCC 25923, E. coli ATCC 25922,
or P._aeruainoæa ATCC 27853 were added to the wells in full
strength ~ueller Hinton broth (BBL 11443) from a mid-log
culture. The inoculum is standardized
spectrophotometrically at 600 nm and is verified by colony
counts. The plates are incubated for 16-20 hours at 37C,
1"~ ' SUI~Sml~TE SJ~E'r
W093/05802 ' PCT/US92/07622
-19- 21I8939
'1 '
and the minimal inhibitory concentration (MIC) for each
peptide i8 determined. Minimal inhibitory concentration is
defined as the lowest concentration of peptide which produc-
es a clear well in the microtiter plate. The minimal
inhibitory concentration of each of (SEQ ID NO:l) through
(SEQ ID NO:3), (SEQ ID NO:5), (SEQ ID NO:6), (SEQ ID NO:9)
through (SEQ ID NO:ll), and (SEQ ID NO:17) is given in Table
I below.
Table I
Minimal Inhibitory Concentration
( ~g/ml)
PeDtide S.aureus P.aeru~inosa E.coli
X-(SEQ ID NO:l)-NH2 16 4,8 4
(SEQ ID NO.:l)-NH2 64,128 8,16 8,16
X-(SEQ ID NO:2)-NH2 16 16 8,16
(SEQ ID NO.:2)-NH2 16 16 8,16
X-(SEQ ID NO:3)-NH2 64 16 8, 16
(SEQ ID NO.:3)-NH2 64 16 8,16,32
X-(SEQ ID NO:S)-NH2 4 8 32
X-(SEQ ID NO:6)-NH2 16 128 8
X-(SEQ ID NO:9)-NH2 16 8 64
X-(SEQ ID NO:10)-N~2 16,32 16 64
(SEQ ID No:Il) >256 2~6 >256
(SEQ ID N0:17)-NH2 >256 32,64 64,128
X-(SEQ ID NO:17)-NH2 256 8,16 32,64
The peptides of the pre~ent invention, whether administered
alone or i~ combination with agents auch as ions ha~ing
pharmalogical properties, antibiotic~, or o~her biologically
active peptides or proteins as hereinabove described, may be
employed in a wide ~ariety of pharmaceutical compositions in
combination with a non-toxic pharmaceutical carrier or vehicle
8uch a~ a filler, non-toxic-bufer, or physiological saline
solution. Such pharmaceutical compositions may be used topically
SUBSmUTE SHEI~
W093/~02 rCT/US92/07622
893 9 -20~
or systemically and may be in any suitable form such as a liquid,
solid, semi-solid, injectable solution, tablet, ointment, lotion,
paste, capsule or the like. The peptide and/or agent as
hereinabove described may also be used in combination with
ad~uvants, protease inhibitorfi, or compatible drugs where such a
combination is ~een to be desirable or advantageous in control-
ling infection caused by harmful microorganisms including proto-
zoa, viruses, parasites, fungi, and the like.
The peptide may be admini6tered to a host in particular an
animal, in an effective antibiotic and/ar anti-tumor and/or
antiviral and/or antimicrobial and/or antispermicidal and/or
antifungal and/or antiparasitic amount, or in an amount effective
to ~timulate wound healing in a host. The peptides may be
administered either alone or in combination with an ion having
pharmacological properties, antibiotic, or ion channel forming
peptide or protein as hereinabove described. When the peptide i5
administered in combination with a ion having pharmacological
properties, the activity of the peptide is potentiated.
When the peptide is administered in combination with an
agent as hereinabove described, it is possible to administer the
peptide and agent in separate forms. For example, the agent may
be~a ~ inistered systemically and the peptide may be administered
topically.
When the peptide is admini~tered topically, it may be
admini~tered in combination with a water-soluble vehicle, said
water-soluble vehicle being in the form of an ointmentj cream,
lotion, paste or the like. Examples of water-soluble vehicles
which may be employed include, but are not limited to, glycol~,
~uch a~ polyethylene glycol, hydroxycellulose, and KY Jelly. The
water-soluble vehicle i~ preferably free of an oily substance.
The peptide may also be employed in combination with a ion
having pharmacological properties, as hereinabove described in
the form of an oral composition for oral hygiene. Such a compo-
sition may be incorporated into a wide variety of compositions
:~
.7,
'J ' ' ~ :
SU8STlT~JTE SHEEt
,~
WOD3/0~2 PCT/US92/07622
- -21- 2118939
and materials used for oral hygiene purposes, which include, but
are not limited to, toothpastes, mouthwashes, tooth gels, and
tooth powder~. Sueh eomposition may thu~ be u~ed to treat or
prevent periodontal disea~e, to prevent or reduce plaque, and/or
to prevent or treat or reduee dental earies. The peptide and
toxie ion may be used to inhibit, prevent, or destroy the growth
of StreDtoeoeeus mutan~, whieh i8 assoeiated with dental carie#
and periodontal dlJease.
Numerous modifieations and variations of the pre~ent inven-
tion are possible in light of the above teaehings; therefore,
within the seope of the appended elaims the invention may be
praeeie-d otherwi-- than ~ part1eulArly d--eribed.
.
. ~ .
.
,~ .
~ ~ ,
, ~ .
,, ~
, ",:,
~ ~ ~ suBsmu~E SHEET
W093/~02 PCT/US92/07622
.2 1 18 9~ 9 -22-
SEQUENOE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT: Maloy, W. Lee
Kari, U. Prasad
Williamæ, Jon I.
- (ii) TITLE.OF INVENTION: Biologically Active Peptide
Compositions.and Uses Therefor
(.iii) NUMBER OF SEQUENCES: 19
.
(iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Carella, Byrn~, Bain, Gilfillan,
Cecchi ~ Stewart
(B) STREET: 6 Becker Farm Road
(C) CITY: Ro~eland
(D) STATE: New Jer~ey
(E) COUNTRY: USA
(F) ZIP: 07068
(v) COMPUTER READABLE FORM:.
(A) MEDIUM TYPE: 3.5 inch diskette
(B~ COMPUTER: IBM PS/2
- (C) OPERATING SYSTEM: PC-DOS
(D) SOFTWARE: DW4.V2
(vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
(vii) PRIOR APPLICATION DATA:
~UBSTtTlJTE SHEE~
:
W093/05802 PCT/VS92/076~2
-23- 2~18939
(A) APPLICATION NUMBER: 07/760,054 :.
(B) FILING DATE: 13-SEP-1991
(viii) ATTORNEY/AGENT INFORMATION:
(A) NAME: Olstein, Elliot M.
(B) REGISTRATION NUMBER: 24,025
(C) REFERENCE/DOCKET NUMBER: 421250-161
.
(ix) TELECOMMUNICATION INFORMATION:
~A) TELEPHONE: 201-994-1700
(B) TELEFAX: 201-994-1744
(2) IN~ORMATION FOR SEQ ID NO:l:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 11 amino acids
(B) ~ E: amino acid
(C) STRA ~ EDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATVRE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at-N-termi-
nus.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:1:
Leu Lys Ly~ Leu Lys Lys Leu Leu Lys Leu
Le~
~2) INFORMATION FOR SEQ ID NO:2:
- (i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 12 amino acids
8UBSTIT~JTE S'~EFr
W093/~02 P~T/US92/07622
2 1 1 893 9 -24-
(B) TYPE: amino acid
(C) STRANDEDNESS:
- ~D) TOPO~OGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(~) OL~R INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:2:
Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys
Leu Leu
(2) INEORMATION FOR SEQ ID NO:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 13 amino acids
5B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE mE peptide
(ix) FEATURE:
(~) OT~ER INFO~MATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nu~.
S~BSTITUTE SHE~T
WO 93/05802 PCI~/US92/07622
-25 ~
2`118939
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:3:
-Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu
5 10
Lys Leu Leu
(2) INFORMATION FOR SEQ ID NO:4:
(i) SEQUENCE CHARACTERISTICS:
(A)`LENGTH: 14 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
,
(ix) FEATURE:
(D) OTEER INFORMATION: May be a C-termlnal amide,
` and/or may be acetylated at N-termi-
nus.
.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:4:
" ~
Lys Lys Leu Leu Lys Lys Leu Lyæ Lys Leu
Leu Lys Leu Leu
(2) INFORMATION FOR SEQ ID NO:5:
(i) SEQUENCE CHARACTERISTICS:
;~; (A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
-
5;
~'. ` : '
SuBsT~ TE sH
W093/~802 PCTJUS92/07622
-26-
(D) Ol~ER INFORMATION: May be a C-terminal
2 ~ ~893 9 amide, and/or may be acetylated at N-termi-
nus.
(ix) SEQUENCE DESCRIPTION: SEQ ID NO:5:
Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Ar~ Arg
(2) INFORMATION FOR SEQ ID NO:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: :
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
( ix) EEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:6:
Lys Lau Lys Lys Leu Leu Lys Lys Leu Lys
Lys Leu Leu Lys Leu Leu
(2) INFORMATION FOR SEQ ID NO:7:
(i) SEQUENCE CHARACTERISTICS:
(A).LENGTH: 15 amino acids
- (B) TYPE: amino acid
( C ) STRANDEDNESS:
(D) TOPOLOGY: linear
SUBSTITUTE SHEET
S ~i ~ 3 ~. '~! ~ . `
W093/05802 PCT/US92/07622
-27- 21I8939
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
- nu~.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:7:
Leu Lys Lys Leu Leu Lys Lys ~eu Ly~ Lys
Leu Leu Ly~ Ly~ Asn
(2) INFORMATION FOR SEQ ID NO:8:
(i) SEQUENCE CHARACTERISTICS:
. (A) LENGTH: 15 amino acid~
: (B) TYPE: amino acid
(C) STRA ~EDNESS:
(D) TOPOLOGY: linear
.
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INEORMATION: May be a C-terminal amide,
and/or may be ace~ylated at N-termi-
nus, Xaa is homoserine.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:8:
Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys
Leu Leu Lys Lys Xaa
. SLJBSTITUTE SHEET
^~
W093/05802 PCT/US92/07622
-28-
2 ~ 3 ~FORMATION FOR SEQ ID NO:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear ~ :
(ii) MOLECULE TYPE: peptide
.
(ix) EEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:9:
Leu Lys Leu Leu Lys Ly~ Leu Leu Lys Lys
:~ 5 10
, ~ .
A~n Lys Lys Leu Leu Lys Ly~Leu
~ : 15
'J';` ~ ( 2) INFORMATION roR SEQ ID NO:10:
~ (i) SEQUENCE.CHARACTERISTICS:
T ~ (A) LENGTH: 18 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
: nu~.
5 i ,,~
,:", ~
SU~SnT~E SH~
~$~
W093/05802 PCT/US92/07622
-29-
2118939
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:lO:
Leu Lys Leu Leu Lys Lys Leu Leu Ly~ Lys
Pro Lys Lys Leu Leu Lys Lys Leu
(2) INFORMATION FOR SEQ ID NO:ll:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 22 amino acid~
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
~,
(ii) MOLECULE TYPE: peptide
( ix) EEAlVRE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
~xi) SEQUENCE DESCRIPTION:SEQ ID NO:ll:
,
: Leu Leu Lys Lys Leu l.y$ Lys Leu Leu Lys
Lys Leu Gln Gly Pro Pro Gln Gly Gln Ser
Pro Gln
(2) INFORMATION FOR SEQ ID NO:12:
(i) SEQUENCE CHARA~TERISTICS:
(A) LENGTH: 20 amino acids
(B) TYPE: amino acid
(C) STRANDE~NESS:
SUBSTITVTE SHEFr
~ ;, iJ. ~ .,t W C
W093/05802 PCT/US92/07622
-30-
2~i~939
- (D) TOPOLOGY: linear
(ii) MOLECULE mE peptide
(ix) FEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
.
(xi) SEQUENCE DESCRI~TION:SEQ I~ NO:12:
Leu Ala Ser Lys Ala Gly Ala Ile Ala Gly
Lys Ile Ala Lys Lys Leu Leu Lys Lys Leu
(2) INFORMATION FOR SEQ ID NO:13:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(iX).E~3ATUR~:
(D) OTHER INFORMATION: May be a C-terminal amida,
and/or may be acetylated at N-termi-
nu~.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:13:
Leu Lys Lys Leu Lys Lys Leu
SUB~;TI~UTE SHl~EI'
W093/058~2 PCT/US92/07622
-31-
2118939
(2) INFORMATION FOR SEQ ID NO:14:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids
(B) m E: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INEORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
'
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:14:
Leu Leu Lys Lys Leu Lys Lys Leu
(2) INFORMATION FOR SEQ ID NO:15:
(i) SEQUENCE C~ARACTERISTICS:
(Aj LENGTH: 9 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
- (ii) MOLECULE TYPE: peptide
(ix) FEATURE:
~D) OTHER INFORMATION: May be a C terminal amide,
and/~r may be acetylated at N-termi-
nus.
SUBSTITUTE S~EET
- ~
2 ~ 32- PCT/US92/076
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:15:
Lys Leu Leu Ly~ Ly~ Leu Lys Lys Leu
(2) INFORMATION EOR SEQ ID NO:16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 10 amino acids
(B) TYPE: amino acid
(C)``STRANDEDNESS:
(D) TOPOLOGY: linear
MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nus.
,
- .
(xi) SEQUENC:E DESCRIPTION:SEQ ID NO:16:
Lys Ly~ Leù~L~u Ly- Ly~ Leu Ly~.Ly~ Leu
r,.~ S 10
(2) INFORMATION EOR SEQ ID NO:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 11 amino acids
(B) TYPE: ami~o acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLEGULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
:
8UBSTITUTE SHEET
~y
_~
WOg3/0~2 PCT/US92/07622
~3~~ 2118939
and/or may be acetylated at N-termi- :
nu~.
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:17:
Leu Lys Lys Leu Leu Lys Ly~ Leu Lys Ly~ Leu
(2) INEORMATION FOR SEQ ID NO:18:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 11 amlno acids
(B) TYrE: amino acid
(C) STRANDEDNESS:
.(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-terml-
nUB .
,~ .
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:18:
Ala Ly~ Ly~ Leu Leu Ly~ Ly~ Léu Lys Lys Leu
(2) INFORMATION FOR SEQ ID NO:l9:
(i) SEQUENCE CHARACTERISTICS:
- . (A) LENGTH: 14 amino acids
(B) TY~E: amino acid
(C) STRANDEDNESS:
(D~ TOPOLOGY: ;linear
(ii) MOLECULE TYPE: peptide
.; .
, ,:
. - :
;,. ~ : ~
$~UBSIITU~E 9HEEr
W093/0~802 PCT/US92/07622
2 i l893 9 ~34~
(ix) FEATURE:
(D) Ol~ER INFORMATION: May be a C-terminal amide,
and/or may be acetylated at N-termi-
nu~. .
(xi) SEQUENCE DESCRIPTION:SEQ ID NO:19: :
Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
Leu Lys Arg
SUBSTIT~TE~
, ~
